The paradigm for this Grand Opportunities (RFA-OD-09-004;NIAAA topic: "Molecular Markers of Alcohol Exposure and Alcohol-induced Tissue Injury") proposal is that alcohol causes tissue injury and organ dysfunction by disrupting the homeostatic interaction between the intestinal microbiome and the human homobiome. We define this homeostatic interaction as the Metabiome and will model the disruption of the Metabiome during Alcoholic Liver Disease using techniques inspired from Systems Biology. We hypothesize that changes in the microbiome pattern and/or function in a sub-group of alcoholics predisposes them to long term medical consequences of alcoholism and this vulnerability is due to the impact of the microbiome on both local (mucosal) and systemic immunity or on local (gut) and systemic lipid metabolism. The goal of this proposal is to identify "Molecular Markers of Alcohol Exposure and Alcohol-induced Tissue Injury" by proposing a series of experiments in alcoholics without and with alcoholic liver disease [ALD] to test the novel hypothesis that microbial initiated changes in systemic immunity and lipid metabolism will determine whether alcohol abuse results in clinically significant tissue injury and hepatic dysfunction. To do this we propose three Specific Aims: Aim 1. To interrogate the Microbiome in sigmoid mucosa and stool of alcoholics with ALD and Control groups. We will use Multitag Pyrosequencing (MTPS), Metatranscriptomic and Metabolomic techniques to identify factors that correlate the changes in microbiota composition and functionality with alcoholic liver disease [ALD]. Aim 2. To assess the local (gut mucosa) and systemic immune state in sigmoid mucosa and serum respectively of alcoholics with ALD and Control groups. We will use immunohistochemistry, RT-PCR, and cytokine assays to determine whether changes in the Immunome (mucosal and global immune state) induced by disruption of "metabiome" correlate with susceptibility to liver injury to alcohol. Aim 3. To assess gut lumen, gut mucosa, and systemic metabolic states of alcoholics with ALD and their Control groups. We will use the Metabolomic techniques to characterize metabolite expression in already stored sigmoid mucosa, lumen (fecal water), serum and urine of patients to determine whether changes in the metabolic states induced by disruption of "metabiome" are correlated with ALD. By identifying the abnormal "Metabiome" components in alcohol-induced liver injury, we will provide a novel approach for the identification of biomarker(s) to identify alcoholics at risk of developing organ dysfunction like liver disease. This grant will support the employment of 8 persons, including two newly hired and excluding the PIs (one of the stated aims of ARRA). PUBLIC HEALTH RELEVANCE: The goal of this proposal is to investigate why only some alcoholics get alcoholic liver disease (ALD) using an approach called Systems Biology which really means looking at many different factors at once to see how they are related and to identify specific new disease 'biomarkers'. Our main hypothesis is that one key reason why some alcoholics get ALD is because chronic alcohol use changes the intestinal bacteria which has long term effects on a person's metabolism and immune system. It can also cause the intestines to be leaky and allow more bad things to enter the blood. In this study we will look at samples from alcoholics with and without liver disease as well as other patient groups and identify the alcohol-related profile of their intestinal bacteria and how it affects their metabolism and immune system. This analysis will allow us to identify new kinds of alcohol- related biomarkers of intestinal bacteria, immune function, and metabolism to help us identify patients at risk for ALD and to help treat ALD patients.